Figure 5. EDEM1 is the downstream molecular target of the LINC00485/miR-581 axis in CRC cells. (A) Schematic diagram of binding sites of the 3’UTR of EDEM1 mRNA and miR-581. (B) Luciferase reporter assay confirming the interaction between the 3’UTR of EDEM1 mRNA and miR-581. (C, D) RIP assay validating the interaction between the 3’UTR of EDEM1 mRNA and miR-581 in (C) FHC cells and (D) LoVo cells. (E) The mRNA level of EDEM1 is significantly lower in 52 CRC tissues than in paired normal tissues. (F, G, H) The (F) mRNA and (G, H) protein levels of EDEM1 in CRC patients with stage I/II or III/IV are significantly lower than in adjacent normal tissues. EDEM1 expression level decreases significantly with the advancing tumor stage. (I) A positive correlation between LINC00485 level and the expression of EDEM1 in 52 CRC tissues. (J) EDEM1 expression is negatively correlated with miR-581 levels in 52 CRC tissues. (K, L) The (K) mRNA and (L) protein levels of are significantly reduced in CRC cells compared with in human normal colorectal epithelial cell lines. Comparison between two groups were assessed using student’s t-test. Multiple comparison was analyzed using the one-way ANOVA with LSD test. Bars were represented as S.D. *P<0.05; **P<0.01; ***P<0.001. WT, wild type; N, adjacent normal tissues; T, tumor tissues; NS, not significant.